# 802: GRACILIBACILLUS MEDIUM


## Main sol. 802

100	g/l	NaCl	
20	g/l	MgSO4 x 7 H2O	
2	g/l	KCl	
0.1	g/l	NaBr	
2	g/l	Yeast extract (Difco)	
2	g/l	Trypticase peptone (BBL)	
12	g/l	Tris-HCl	
2	ml	**Trace metals solution**	
998	ml	Distilled water	

1. Adjust pH to 7.8 (with NaOH) before autoclaving. To the sterile medium, from sterile stock solutions, add 10 ml KH2PO4 (50 g/l), 5 ml CaCl2 (100 g/l), 2 ml FeCl2/MnCl2 (FeCl2 x 4 H2O, 20 g/l and MnCl2 x 4 H2O, 20 g/l), and glucose to a final concentration of 2 g/l.

10	ml	KH2PO4 (50 g/l)	
5	ml	CaCl2 (100 g/l)	
2	ml	**FeCl2/MnCl2 solution**	
2	g	Glucose	


## Trace metals solution

10	ml	HCl (32%)	
2	g	FeCl2 x 4 H2O	
250	mg	CoCl2 x 6 H2O	
100	mg	MnCl2 x 4 H2O	
70	mg	ZnCl2	
6	mg	H3BO3	
40	mg	Na2MoO4 x 2 H2O	
70	mg	NiCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
60	mg	AlCl3 x 6 H2O	
6	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve salts in the HCl in the oder given above, then make up the solution to 1 litre with distilled water.


## FeCl2/MnCl2 solution

20	g	FeCl2 x 4 H2O	
20	g	MnCl2 x 4 H2O